KC-4920

Jurakt-NFAT-Luc2-CD32b Cell Line

54347

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Background of Jurakt-NFAT-Luc2-CD32b Cell Line

The protein encoded by this gene is a low affinity receptor for the Fc region of immunoglobulin gamma complexes. The encoded protein is involved in the phagocytosis of immune complexes and in the regulation of antibody production by B-cells. Variations in this gene may increase susceptibilty to systemic lupus erythematosus (SLE). Several transcript variants encoding different isoforms have been found for this gene.FCGR2B (Fc Gamma Receptor IIb, CD32b) is a Protein Coding gene. Diseases associated with FCGR2B include Systemic Lupus Erythematosus and Malaria. Among its related pathways are Fc-GammaR Pathway and Immune response Fc epsilon RI pathway.

Specifications

Catalog Number	KC-4920
Cell Line Name	Jurakt-NFAT-Luc2-CD32b Cell Line
Clone Number	2B2
Host Cell Line	Jurakt-NFAT-Luc2
Description	Stable Jurkat cell line expressing exogenous luciferase under the control of NFAT responsive element and CD32b fusion sequence.
Quantity	One vial of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 300μg/mL Hygromycin B + 1μg/mL Puromycin
Selection Marker	Hygromycin B, Puromycin
Morphology	Lymphoblast
Subculture	Split saturated culture at a ratio of 1:2~1:4 every 2~3 days; seed out at about 1 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

Jurakt-NFAT-Luc2-CD32b Cell Line was generated using a lentiviral vector expressing the luciferase sequence.

Characterization

Figure 1: Jurkat-NFAT-Luc2-CD32b cells and Raji cells were seeded into 96-well plates, treated with Rituximab(Cat# KB-1122, Kyinno) in different concentrations for 16 hours, and then read out using Bright-Glo Detection System.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS, 300µg/mL Hygromycin B, 1μg/mL Puromycin) in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture at a ratio of 1:2~1:4 every 2~3 days; seed out at about 1 × 10⁵ cells/mL

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage.

References

1. Wang Y, Sugita N, Kikuchi A, Iwanaga R, Hirano E, Shimada Y, Sasahara J, Tanaka K, Yoshie H. FcγRIIB-nt645+25A/G gene polymorphism and periodontitis in Japanese women with preeclampsia. Int J Immunogenet. 2012 Dec;39(6):492-500. doi: 10.1111/j.1744-313X.2012.01124.x. Epub 2012 May 18. PMID: 22594540.
2. Aitman TJ, Cooper LD, Norsworthy PJ, Wahid FN, Gray JK, Curtis BR, McKeigue PM, Kwiatkowski D, Greenwood BM, Snow RW, Hill AV, Scott J. Malaria susceptibility and CD36 mutation. Nature. 2000 Jun 29;405(6790):1015-6. doi: 10.1038/35016636. PMID: 10890433.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.